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. 1991 Feb;11(2):1099–1106. doi: 10.1128/mcb.11.2.1099

Identification of regulatory sequences and protein-binding sites in the liver-type promoter of a gene encoding 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase.

F P Lemaigre 1, S M Durviaux 1, G G Rousseau 1
PMCID: PMC359787  PMID: 1846662

Abstract

The liver-type and muscle-type isozymes of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase are encoded by one gene that uses two alternative promoters. We have identified cis-acting sequences and protein-binding sites on the liver-type promoter. Transfection assays with deleted promoters showed that maximal promoter activity is contained within 360 bp upstream of the cap site. DNase I footprinting experiments with liver and spleen nuclear extracts and with purified proteins revealed several protein-binding sites in this region. These included four binding sites for nuclear factor I, one site that contains an octamer consensus but showed a liver-specific footprint pattern, two liver-specific protein-binding sites, and one poly(dG)-containing binding site. Transfection of cells of hepatic origin suggested that all these sites except one are involved in transcriptional regulation. The region between -360 and -2663 contained an element that functioned as a silencer in a nonhepatic cell line. We conclude that in liver transcription from the liver-type promoter of the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene is controlled by ubiquitous and tissue-specific factors and involves activating and derepressing mechanisms.

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Selected References

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  1. Beato M. Gene regulation by steroid hormones. Cell. 1989 Feb 10;56(3):335–344. doi: 10.1016/0092-8674(89)90237-7. [DOI] [PubMed] [Google Scholar]
  2. Clark S. P., Lewis C. D., Felsenfeld G. Properties of BGP1, a poly(dG)-binding protein from chicken erythrocytes. Nucleic Acids Res. 1990 Sep 11;18(17):5119–5126. doi: 10.1093/nar/18.17.5119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Colosia A. D., Marker A. J., Lange A. J., el-Maghrabi M. R., Granner D. K., Tauler A., Pilkis J., Pilkis S. J. Induction of rat liver 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase mRNA by refeeding and insulin. J Biol Chem. 1988 Dec 15;263(35):18669–18677. [PubMed] [Google Scholar]
  4. Courtois S. J., Lafontaine D. A., Lemaigre F. P., Durviaux S. M., Rousseau G. G. Nuclear factor-I and activator protein-2 bind in a mutually exclusive way to overlapping promoter sequences and trans-activate the human growth hormone gene. Nucleic Acids Res. 1990 Jan 11;18(1):57–64. doi: 10.1093/nar/18.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crepin K. M., Darville M. I., Hue L., Rousseau G. G. Characterization of distinct mRNAs coding for putative isozymes of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. Eur J Biochem. 1989 Aug 1;183(2):433–440. doi: 10.1111/j.1432-1033.1989.tb14946.x. [DOI] [PubMed] [Google Scholar]
  6. Crossley M., Brownlee G. G. Disruption of a C/EBP binding site in the factor IX promoter is associated with haemophilia B. Nature. 1990 May 31;345(6274):444–446. doi: 10.1038/345444a0. [DOI] [PubMed] [Google Scholar]
  7. Darville M. I., Crepin K. M., Hue L., Rousseau G. G. 5' flanking sequence and structure of a gene encoding rat 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6543–6547. doi: 10.1073/pnas.86.17.6543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Decaux J. F., Antoine B., Kahn A. Regulation of the expression of the L-type pyruvate kinase gene in adult rat hepatocytes in primary culture. J Biol Chem. 1989 Jul 15;264(20):11584–11590. [PubMed] [Google Scholar]
  9. Gorski K., Carneiro M., Schibler U. Tissue-specific in vitro transcription from the mouse albumin promoter. Cell. 1986 Dec 5;47(5):767–776. doi: 10.1016/0092-8674(86)90519-2. [DOI] [PubMed] [Google Scholar]
  10. Gounari F., De Francesco R., Schmitt J., van der Vliet P., Cortese R., Stunnenberg H. Amino-terminal domain of NF1 binds to DNA as a dimer and activates adenovirus DNA replication. EMBO J. 1990 Feb;9(2):559–566. doi: 10.1002/j.1460-2075.1990.tb08143.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hollon T., Yoshimura F. K. Variation in enzymatic transient gene expression assays. Anal Biochem. 1989 Nov 1;182(2):411–418. doi: 10.1016/0003-2697(89)90616-7. [DOI] [PubMed] [Google Scholar]
  12. Hue L., Rider M. H. Role of fructose 2,6-bisphosphate in the control of glycolysis in mammalian tissues. Biochem J. 1987 Jul 15;245(2):313–324. doi: 10.1042/bj2450313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lenardo M. J., Staudt L., Robbins P., Kuang A., Mulligan R. C., Baltimore D. Repression of the IgH enhancer in teratocarcinoma cells associated with a novel octamer factor. Science. 1989 Jan 27;243(4890):544–546. doi: 10.1126/science.2536195. [DOI] [PubMed] [Google Scholar]
  14. Lewis C. D., Clark S. P., Felsenfeld G., Gould H. An erythrocyte-specific protein that binds to the poly(dG) region of the chicken beta-globin gene promoter. Genes Dev. 1988 Jul;2(7):863–873. doi: 10.1101/gad.2.7.863. [DOI] [PubMed] [Google Scholar]
  15. Marker A. J., Colosia A. D., Tauler A., Solomon D. H., Cayre Y., Lange A. J., el-Maghrabi M. R., Pilkis S. J. Glucocorticoid regulation of hepatic 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene expression. J Biol Chem. 1989 Apr 25;264(12):7000–7004. [PubMed] [Google Scholar]
  16. Mermod N., O'Neill E. A., Kelly T. J., Tjian R. The proline-rich transcriptional activator of CTF/NF-I is distinct from the replication and DNA binding domain. Cell. 1989 Aug 25;58(4):741–753. doi: 10.1016/0092-8674(89)90108-6. [DOI] [PubMed] [Google Scholar]
  17. Mueller C. R., Maire P., Schibler U. DBP, a liver-enriched transcriptional activator, is expressed late in ontogeny and its tissue specificity is determined posttranscriptionally. Cell. 1990 Apr 20;61(2):279–291. doi: 10.1016/0092-8674(90)90808-r. [DOI] [PubMed] [Google Scholar]
  18. Nguyen V. T., Morange M., Bensaude O. Firefly luciferase luminescence assays using scintillation counters for quantitation in transfected mammalian cells. Anal Biochem. 1988 Jun;171(2):404–408. doi: 10.1016/0003-2697(88)90505-2. [DOI] [PubMed] [Google Scholar]
  19. Nordeen S. K. Luciferase reporter gene vectors for analysis of promoters and enhancers. Biotechniques. 1988 May;6(5):454–458. [PubMed] [Google Scholar]
  20. Ochoa A., Brunel F., Mendelzon D., Cohen G. N., Zakin M. M. Different liver nuclear proteins binds to similar DNA sequences in the 5' flanking regions of three hepatic genes. Nucleic Acids Res. 1989 Jan 11;17(1):119–133. doi: 10.1093/nar/17.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Paonessa G., Gounari F., Frank R., Cortese R. Purification of a NF1-like DNA-binding protein from rat liver and cloning of the corresponding cDNA. EMBO J. 1988 Oct;7(10):3115–3123. doi: 10.1002/j.1460-2075.1988.tb03178.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rosenfeld P. J., Kelly T. J. Purification of nuclear factor I by DNA recognition site affinity chromatography. J Biol Chem. 1986 Jan 25;261(3):1398–1408. [PubMed] [Google Scholar]
  23. Rupp R. A., Kruse U., Multhaup G., Göbel U., Beyreuther K., Sippel A. E. Chicken NFI/TGGCA proteins are encoded by at least three independent genes: NFI-A, NFI-B and NFI-C with homologues in mammalian genomes. Nucleic Acids Res. 1990 May 11;18(9):2607–2616. doi: 10.1093/nar/18.9.2607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rupp R. A., Sippel A. E. Chicken liver TGGCA protein purified by preparative mobility shift electrophoresis (PMSE) shows a 36.8 to 29.8 kd microheterogeneity. Nucleic Acids Res. 1987 Dec 10;15(23):9707–9726. doi: 10.1093/nar/15.23.9707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Santoro C., Mermod N., Andrews P. C., Tjian R. A family of human CCAAT-box-binding proteins active in transcription and DNA replication: cloning and expression of multiple cDNAs. Nature. 1988 Jul 21;334(6179):218–224. doi: 10.1038/334218a0. [DOI] [PubMed] [Google Scholar]
  26. Schaffner W. How do different transcription factors binding the same DNA sequence sort out their jobs? Trends Genet. 1989 Feb;5(2):37–39. doi: 10.1016/0168-9525(89)90017-6. [DOI] [PubMed] [Google Scholar]
  27. Schöler H. R., Dressler G. R., Balling R., Rohdewohld H., Gruss P. Oct-4: a germline-specific transcription factor mapping to the mouse t-complex. EMBO J. 1990 Jul;9(7):2185–2195. doi: 10.1002/j.1460-2075.1990.tb07388.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schöler H. R., Hatzopoulos A. K., Balling R., Suzuki N., Gruss P. A family of octamer-specific proteins present during mouse embryogenesis: evidence for germline-specific expression of an Oct factor. EMBO J. 1989 Sep;8(9):2543–2550. doi: 10.1002/j.1460-2075.1989.tb08392.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wall S. R., van den Hove M. F., Crepin K. M., Hue L., Rousseau G. G. Thyroid hormone stimulates expression of 6-phosphofructo-2-kinase in rat liver. FEBS Lett. 1989 Nov 6;257(2):211–214. doi: 10.1016/0014-5793(89)81536-4. [DOI] [PubMed] [Google Scholar]
  30. Wigler M., Pellicer A., Silverstein S., Axel R. Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor. Cell. 1978 Jul;14(3):725–731. doi: 10.1016/0092-8674(78)90254-4. [DOI] [PubMed] [Google Scholar]

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